Recombinant Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401] (ab201015)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR19401] to ERK1 (phospho T202) + ERK2 (phospho T185)
- Suitable for: ICC/IF, IP, IHC-P, WB, Dot blot
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401] -
Description
Rabbit monoclonal [EPR19401] to ERK1 (phospho T202) + ERK2 (phospho T185) -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, IP, IHC-P, WB, Dot blotmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Jurkat, treated with 200 ng/ml PMA for 30 minutes whole cell lysate; NIH/3T3, untreated and treated with 50 ng/ml PDGF for 40 minutes whole cell lysates; PC-12, untreated and treated with 200 ng/ml NGF for 4 days whole cell lysates. IHC-P: Human breast and glioma tissues. IP: NIH/3T3 treated with 50ng/ml PDGF for 40min whole cell lysate; PC-12 treated with 100ng/ml NGF for 10min whole cell lysate. ICC/IF:Jurkat
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 0.05% BSA, 40% Glycerol -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR19401 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab201015 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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ICC/IF | (3) |
Use at an assay dependent concentration.
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IP |
1/30.
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IHC-P |
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IHC is recommended for human only. |
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WB | (1) |
1/1000. Detects a band of approximately 44, 42 kDa (predicted molecular weight: 41 kDa).
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Dot blot |
1/1000.
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Notes |
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ICC/IF
Use at an assay dependent concentration. |
IP
1/30. |
IHC-P
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. IHC is recommended for human only. |
WB
1/1000. Detects a band of approximately 44, 42 kDa (predicted molecular weight: 41 kDa). |
Dot blot
1/1000. |
Target
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Cellular localization
ERK2: Nucleus. -
Database links
- Entrez Gene: 5594 Human
- Entrez Gene: 5595 Human
- Entrez Gene: 26413 Mouse
- Entrez Gene: 26417 Mouse
- Entrez Gene: 116590 Rat
- Entrez Gene: 50689 Rat
- Omim: 176948 Human
- Omim: 601795 Human
see all
Images
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All lanes : Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401] (ab201015) at 1/1000 dilution
Lane 1 : Untreated Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
Lane 2 : Jurkat (Human T cell leukemia cell line from peripheral blood) treated with 200 ng/ml PMA for 30 minutes whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 41 kDa
Observed band size: 42,44 kDa why is the actual band size different from the predicted?
Exposure time: 15 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Jurkat (Human T cell leukemia cell line from peripheral blood) cells labeling ERK1 (phospho T202) and ERK2 (phospho T185)
ERK1 (phospho T202) + ERK2 (phospho T185) with ab201015 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing staining on M phase cells (PMID:26529125). After PMA treatment (200 ng/ml, 30min), the staining was increased, and LP treatment decreased the PMA induced staining.
The nuclear counter stain is DAPI (blue).
Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (Alexa Fluor® 594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
The negative controls are as follows:
-ve control 1: ab201015 at 1/500 dilution followed by ab150120 (Alexa Fluor® 594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor® 488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution. -
Immunohistochemical analysis of paraffin-embedded Human glioma tissue labeling ERK2 (phospho T185) with ab201015 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear with weak cytoplasm staining on Human glioma is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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All lanes : Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401] (ab201015) at 1/1000 dilution
Lane 1 : Untreated NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate
Lane 2 : NIH/3T3 (Mouse embryonic fibroblast cell line) treated with 50 ng/ml PDGF for 40 minutes whole cell lysate
Lane 3 : Untreated PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
Lane 4 : PC-12 (Rat adrenal gland pheochromocytoma cell line) treated with 200 ng/ml NGF for 4 days whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 41 kDa
Observed band size: 42,44 kDa why is the actual band size different from the predicted?
Exposure time: 10 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
The induction conditions refer to PMID:12454035; PMID:17026715; PMID:22206868.
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Dot blot analysis of ERK2 (phospho T185) labeled with ab201015 at 1/1000 dilution.
Lane 1: ERK2 (pT185) phospho peptide: DHTGFLT(p)EYVATR aa179-191 peptide.
Lane 2: ERK2 Non-phospho peptide: DHTGFLTEYVATR aa179-191 peptide.
Lane 3: ERK2 (pY187) phospho peptide: DHTGFLTEY(p)VATR aa179-191 peptide.Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated (ab97051) at 1/100000 dilution was used as secondary antibody.
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.
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Immunohistochemical analysis of paraffin-embedded Human breast tissue labeling ERK2 (phospho T185) with ab201015 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on Human breast is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ERK2 (phospho T185) was immunoprecipitated from 0.35 mg of NIH/3T3 (Mouse embryonic fibroblast cell line) treated with 50ng/ml PDGF for 40min whole cell lysate with ab201015 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab201015 at 1/1000 dilution. VeriBlot for IP Detection Reaction (HRP) (ab131366), was used for detection at 1/1000 dilution.
Lane 1: NIH/3T3 treated with 50ng/ml PDGF for 40min whole cell lysate, 10µg (Input).
Lane 2: ab201015 IP in NIH/3T3 treated with 50ng/ml PDGF for 40min whole cell lysate.
Lane 3: Rabbit IgG,monoclonal [EPR25A]-Isotype Control (ab172730) instead of ab201015 in NIH/3T3 treated with 50ng/ml PDGF for 40min whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes.
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ERK2 (phospho T185) was immunoprecipitated from 0.35 mg of PC-12 (Rat adrenal gland pheochromocytoma cell line) treated with 100ng/ml NGF for 10min whole cell lysate with ab201015 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab201015 at 1/1000 dilution. VeriBlot for IP Detection Reaction (HRP) (ab131366), was used for detection at 1/1000 dilution.
Lane 1: PC-12 treated with 100ng/ml NGF for 10min whole cell lysate, 10µg (Input).
Lane 2: ab201015 IP in PC-12 treated with 100ng/ml NGF for 10min whole cell lysate.
Lane 3: Rabbit IgG,monoclonal [EPR25A]-Isotype Control (ab172730) instead of ab201015 in PC-12 treated with 100ng/ml NGF for 10min whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (157)
ab201015 has been referenced in 157 publications.
- Sun B et al. Irisin reduces bone fracture by facilitating osteogenesis and antagonizing TGF-β/Smad signaling in a growing mouse model of osteogenesis imperfecta. J Orthop Translat 38:175-189 (2023). PubMed: 36439629
- Yin R et al. Corin is regulated by circ-0012397/miR-200a-3p and inhibits the oxygen-glucose deprivation-induced apoptosis of SHSY5Y neuroblastoma cells. Ann Transl Med 10:1242 (2022). PubMed: 36544654
- Li S et al. Puerarin improves diabetic wound healing via regulation of macrophage M2 polarization phenotype. Burns Trauma 10:tkac046 (2022). PubMed: 36568527
- Dai Z et al. Dipeptidyl peptidase-4 is associated with myogenesis in patients with adolescent idiopathic scoliosis possibly via mediation of insulin sensitivity. J Orthop Surg Res 17:82 (2022). PubMed: 35139864
- Tang HY et al. PDGFRβ modulates aerobic glycolysis in osteosarcoma HOS cells via the PI3K/AKT/mTOR/c-Myc pathway. Biochem Cell Biol 100:75-84 (2022). PubMed: 34678088